• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.277-281
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• 1995

The present study was performed to investigate the effect of calcium ion on the mesophyll protoplast culture of Arabidopsis thaliana. The mesophyll protoplase were isolated and cultured on an IMH medium supplemented with CaCl$_2$of various concentrations. When the protoplasts were cultured on the medium containing 0 to 12.5 mM CaCl$_2$, extreme vacuolization occurred without cell division. When the protoplasts were cultured with higher levels of CaCl$_2$ up to 50 mM, vacuolization decreased dose-dependent): and the number of plasma-rich cells increased. Cell division was induced when the protoplast were cultured on the medium with CaCl$_2$ higher than 25 mM. The highest plating efficiency (5-6%) was obtained with 50 mM CaCl$_2$. However the plating efficiency was markedly inhibited by 100mM CaCl$_2$ or above. These resole suggest that the relatively high concentration (50 mM) of calcium ion may be required for the culture of protoplasts.

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.339-343
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• 1995

This study was performed to investigate the effect of iron ion on the mesophyll protoplast culture of Arabidopsis thaliana. Mesophyll protoplasts were isolated and cultured in a modified IMH medium supplemented with various concentrations of Fe-EDTA. Relatively low concentration of Fe-EDTA (<0.02 mM) induced the low level (4.8%) of cell division. In addition the cell division and microcallus growth were dose-dependently stimulated by 0 to 1 mM of Fe-EDTA. In 0.5 to 1 mM concentration range of Fe-EDTA, microcolonies were readily formed and the plating deficiency (8.5%) also showed maximal rate. However more than 1 mM of Fe-EDTA inhibited the initial growth of protoplase. The overall results suggest that Fe2+ion concentration plays an important role at the early developmental stage of protoplast regeneration.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.238.1-238
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• 1998

No Abstract, See Full Text

• Molecules and Cells
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• v.39 no.10
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• pp.768-775
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• 2016

The Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.

• Journal of Plant Biotechnology
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• v.47 no.4
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• pp.283-288
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• 2020

Although the evolution of Arabidopsis thaliana and humans diverged approximately 1.6 billion years ago, recent studies have demonstrated that protein function and cellular processes involved in disease response remain remarkably conserved. Particularly, γ-secretase, a multisubunit protein complex that participates in intramembrane proteolysis (RIP) regulation, is also known to mediate the cleavage of more than 80 substrates including the amyloid precursor protein (APP) and the Notch receptor. Although the genes (PS1/2, APH-1, PEN-2, and NCT) coding for the γ-secretase complex components are present in plant genomes, their function remains largely uncharacterized. Given that the deposition of 42 amino acid long amyloid-β peptides (hAβ₄₂) is thought to be one of the main causes of Alzheimer's disease, we aimed to examine the physiological effects of hAβ₄₂ peptides on plants. Interestingly, we found that Arabidopsis protoplast death increased after 24 h of exposure to 3 or 5 µM hAβ₄₂ peptides. Furthermore, transgenic Arabidopsis plants overexpressing the hAβ₄₂ gene exhibited changes in primary root length and silique phyllotaxy. Taken together, our results demonstrate that hAβ₄₂ peptides, a metazoan protein, significantly affect Arabidopsis protoplast viability and plant morphology.

• Korean Journal of Plant Tissue Culture
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• v.26 no.2
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• pp.127-132
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• 1999

Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.